Abstract: 【Objective】 The study was to improve the multiplication times and quality of peony callus to reduce the browning in the process of proliferation and provide a way for the proliferation of peony callus. 【Methods】 Different additive combinations and concentrations, different pre-culture time and other methods were explored with peony cotyledon and hypocotyl callus as materials. 【Results】 The best medium combinations for the multiplication of peony hypocotyl callus and cotyledon callus are PVP 1.0 mg/L+IAA 0.2 mg/L+TDZ 1.0 mg/L+GA3 1.0 mg/L and PVP 2.0 mg/L+IAA 0.5 mg/L+TDZ 0.5 mg/L+GA3 1.0 mg/L, and the proliferation times are 5.75 and 5.50, respectively. The use of explants that have been pre-cultured for 30d can effectively increase the proliferation times and reduce browning. GA3 1.0 mg/L is beneficial to inhibit browning during callus proliferation and promoted callus proliferation. The callus in the form of light yellowish green floccule on the surface with spongy texture is not prone to brown during subculture multiplication, with the volume proliferation changing significantly in the middle and late stages. 【Conclusion】 The addition of 1.0 mg/L of GA3 and the use of the callus that has been pre-cultured for 30d in the process of peony callus proliferation can achieve higher proliferation times and effectively inhibit browning. Light yellowish green and spongy callus is a sound condition of callus proliferation.